Humans ; MicroRNAs ; Muscle, Smooth, Vascular ; Myocytes, Smooth Muscle ; Phenotype

LPS stimulation of macrophages production of IFN-beta plays a key role in innate immunity defending the microbial invasion. In this study, the effect of S632A3 promoting LPS-induced IFN-beta production and the underlying mechanism were investigated, mRNA level was measured by real-time PCR, cytokine production was determined by ELISA, GSK-3beta activity was investigated by kinase assay, protein phosphorylation and expression were evaluated by Western blotting. The results revealed that S632A3 significantly augmented IFN-beta production by LPS-stimulated macrophages. S632A3 inhibition of the activation of GSK-3beta, reduced the threonine 239 phosphorylation of transcription factor c-Jun but increased the total level of c-Jun in LPS-stimulated macrophages. Moreover, small interfering RNA-mediated knockdown of c-Jun level abrogated the ability of S632A3 to augment IFN-beta. The study thus demonstrates S632A3 being a new anti-inflammation lead compound and provides a molecular mechanism by which S632A3 promoted LPS-induced IFN-beta production in macrophages through inhibiting the activation of GSK-3beta.

Objective To estimate the exposure level of PM2.5,CO and O_3 in the elders in a community in Beijing.Methods The concentrations of PM2.5,CO and O_3 in 10 main activity sites of the elders were measured and 24 h time-activity data of 30 elders was collected by recording of activity log paper,Nov.28,2007—Jan.17,2008.Results The 24 h average exposure concentrations of PM2.5,CO and O_3 for the elders were(146.54?6.60)?g/m~3,(2.67?0.18)mg/m~3 and(32.30?2.79)?g/m~3, respectively,there was no significant difference in the exposure level of PM2.5 and O_3 between male and female elders except CO (P

Objective To investigate safety managements of surgical instruments employed for the minimally invasive surgery through link management so as to guarantee surgery safety. Methods All procedure managements for surgical instruments were performed, such as distribution after sterilization, surgical use,takeover after the surgery for 1 910 cases from July to December 2009 in our hospital. All the surgical instruments involved in the surgery included thoracoscopes, laparoscope, uteroscope, ventriculoscope and surgical robots. We organized management training of surgical instruments for medical staff, filed out management procedures,enhanced their consciousness of safety management, controlled the key link for quality control, performed gap management, handled the existing problems and improved management quality. Results The safety procedure management of surgical instruments was performed for the 1 910 cases, which insured that the surgery could go smoothly. Conclusions Safety management measures are established on the basis of the information traced from surgical use, takeover after the surgery, cleaning, sorting, inspection for surgical instruments so that oversight over its management could be effectively avoided, making its clinical effects more obvious.

In the past few decades, our understanding of platelets has made great progress. Platelets play an unexpected central role in cancer and greatly affect the behavior of cancer cells. At the same time, the physiology and phenotype of platelets are also affected by cancer cells. Therefore, platelet-based tumor targeted therapy strategies have attracted the attention of researchers, but the limitations of their application require more attention. In this paper, the strategies of platelet-based tumor targeted therapy are summarized, and the strategies of platelet mimicking nanocarrier delivery, platelet hitch riding, platelet membrane coating biomimetic and engineered platelet targeting are mainly introduced. The easy activation, hard storage and unknown functional and phenotypic changes of platelets were discussed. At the same time, the strategy of platelet-based targeted tumor therapy is reviewed from theoretical basis and practical application. The development potential of platelets in the field of tumor diagnosis and treatment is discussed, which will provide some theoretical reference for the study of platelet-related tumor diagnosis and targeted therapy.

Objective studyring the proven and probable invasive pulmonary aspergillosis(IPA) eases of some hospitals in Shanghai to provide evidence fur the improvement of IPA clinical diagnosis and therapy.Methotis Fortv-nine IPA cases were retrospectively analyzed for demography data,host tactors,underlying conditions.chest CT,microorganism and histopathology examination,as well as therapy and clinical outcome.ResultsOf 49 subjects including 19(38.8%)proven and 30(61.2%)probable IPA,3 pailents(6.1%)had no host factors,25 patients(51.0%)had IPA associated host factors and underlying conditions.while 21 patients(42.9%)had uncertained fundamental diseases.Chest CT evaluation demonstrated that radiological lesions include nodules in 29 patients,patching in 15,mass in 12,consolidation in 10.cavitation in 34,Halo sign in 19,air bronchogram in 18,crescentic sign in 6,bilateral in 33 and multifocal lesions in 38.The yielding rate of fungus culture in sputum was 26.5%(13/49),and in bronchoalveolar lavage fluid was 66.7%(10/15).Eleven of thirty-six patients(30.6%)had positive results of serum galactomannan antigen tests.Nineteen of twenty-one patients(90.5%)were proven as IPA by lung histologic examinations.Aspergillus fumigatus was the most common pathogen 81.0%(17/21).The responding rate to initial anti-fugus therapy wag 50%(21/42).Conclusion Our study suggests that in IPA patients,bilateral,muhifocal and nodular lesion could be the most common radiological characteristic,while Halo and crescentic sign occar occasionally.Invasive technologies are more valuable to IPA diagnosis.

AIM To investigate the protective effect of serum containing Shenshuaikang Enema Liquid (Rhei Radix et Rhizoma,Salviae miltiorrhizae Radix et Rhizoma,Carthami Flos,Astragali Radix) on HK-2 cells injured by hypoxia/reoxygenation (H/R) and its effect on Wnt/β-catenin pathway.MEHTODS Six New Zealand white rabbits randomly divided into control group,Shenshuaikang high dose group and PBS group (n =2) were treated accordingly for serum collection after 3 days' enema.The HK-2 cells injured by hypoxia/reoxygenation were then administered with serum of rabbits from the control group,PBS group,Shenshuaikang groups (high dose,middle dose and low dose groups due to the differently diluted concentrations) respectively.The H/R damage was determined by ROS fluorescence probe,the cellular damage/apoptosis were analyzed by CFSE/PI method and flow cytometry combined with Annexin V-FITC/PI,and the investigation on effects of drugs on expression of Wnt4 mRNA and β-catenin mRNA were accomplished by fluorescence quantitative PCR.RESULTS The fluorescence intensity of intracellular ROS expression was significantly increased after modeling.CFSE/PI double staining showed that the variant Shenshuaikang dose groups displayed obvious proportional mortality superiority to either the control group or PBS group,and the high dose group achieved the lowest mortality.Annexin V-FITC/PI and flow cytometry showed that,at 12 h,compared with the control group (45.6 ± 2.2)％ and PBS group (41.6 ±0.7) ％,Shenshuaikang groups [low dose group (14.8 ± 0.3) ％,middle dose group (10.3 ± 0.6) ％,high dose group (12.9 ± 0.9)％] obviously inhibited apoptosis/death.Shenshuaikang Enema Liquid medicated serum demonstrated its significant effect on the increase of the Wnt4 mRNA expression and a dual-directional regulation on the expression of β-caterin mRNA by quantitative PCR (P ＜ 0.05).CONCLUSION Inhibition of the apoptosis/death of HK-2 cells with hypoxia/reoxygenation injury due to the serum containing Shenshuaikang Enema Liquid suggests the agent's influence on the activation of Wnt/β-catenin pathway.

Homo sapiens longevity assurance homologue 2 (LASS2) is a novel gene isolated from a human liver cDNA library by our laboratory, and it is a human homologue of the yeast longevity assurance gene LAG1 (Saccharomyces cerevisiae longevity assurance gene). According to our previous results, LASS2 could interact with subunit c of vacuolar type H(+)-ATPase (V-ATPase), and the overexpression of LASS2 could inhibit the cell growth of a human hepatocellular carcinoma (HCC) cell line, SMMC-7721. In order to understand the role of the interaction between LASS2 and V-ATPase in HCC cell growth, we transiently transfected plasmid pCMV-HA2-LASS2 into HCCLM3, a HCC cell line without the significant expression of endogenous LASS2. The pH-sensitive fluorescence probes, BCECF and BCECF-AM, were used to measure the intracellular and extracellular H(+) concentrations of HCCLM3 cells respectively. The effect of LASS2 gene on apoptosis was evaluated with Annexin-V/FITC and propidium iodide (PI) by flow cytometry. Western blot was used to detect cytochrome c (Cyt c) in the cytosol and mitochondria, as well as pro-caspase-3 in cytosol. The results showed that the cell growth of LASS2-transfected HCCLM3 cells was significantly inhibited compared with that of the mock control. LASS2 transfection increased intracellular H(+) concentration of HCCLM3 cells, while decreased extracellular H(+) concentration. Moreover, LASS2 transfection significantly enhanced the apoptosis of HCCLM3 cells. In LASS2-transfected cells, the amounts of Cyt c increased in the cytosol, while decreased in the mitochondria. Meanwhile, the expression of pro-caspase-3 in the cytosolic extracts was decreased. These results implicate that LASS2 gene might increase intracellular H(+) of HCC cells via the interaction with V-ATPase, thereby inducing cell apoptosis through mitochondrial pathway.
Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Liver Neoplasms ; pathology ; Membrane Proteins ; metabolism ; RNA, Small Interfering ; Sphingosine N-Acyltransferase ; metabolism ; Transfection ; Tumor Suppressor Proteins ; metabolism ; Vacuolar Proton-Translocating ATPases ; metabolism

OBJECTIVETo explore the mechanism of anisodamine in treating infectious shock through studying effect of anisodamine on endotoxin lipopolysaccharide (LPS) induced expression of tissue factor (TF) and plasminogen activator inhibitor type 1 (PAI-1) in vascular endothelial cells (EC).

METHODSHuman umbilical vein endothelial cells (HUVEC) were cultured by trypsin digestion method. PAI-1 was measured in the conditioned medium of HUVEC by a specific enzyme-linked immunosorbent assay (ELISA), whereas TF activity was measured in the lysates of these cells by using a single step clotting assay. Specific mRNA expressions were determined by Northern blotting. In order to evaluate a possible contribution of the nuclear factor-kappa B (NF-kappa B) pathway on the transductive effects observed, electrophoretic mobility shift assays (EMSA) were performed using nuclear extracts from HUVEC and NF-kappa B binding oligonucleotides.

RESULTSLPS could significantly strengthen the expression of HUVEC PAI-1 protein and TF activity and its mRNA, this effect of LPS could be markedly weakened after adding Anisodamine dose-dependently. Anisodamine could also completely block the LPS induced NF-kappa B DNA binding activity in nuclear extracts from HUVEC.

CONCLUSIONThe possible mechanism of anisodamine in treating infectious shock may be through antagonizing LPS induced HUVEC TF and PAI-1 expression, and the antagonism might be, at least partially, transduced by path of NF-kappa B.

Cells, Cultured ; Culture Media, Conditioned ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; cytology ; metabolism ; Humans ; NF-kappa B ; metabolism ; Plasminogen Activator Inhibitor 1 ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Solanaceous Alkaloids ; pharmacology ; Thromboplastin ; biosynthesis ; genetics ; Umbilical Veins ; cytology

OBJECTIVETo investigate the changes in plasma levels of vascular endothelial growth factor (VEGF) and stromal cell-derived factor-1 (SDF-1) and in peripheral CD34(+) cells in patients with acute myocardial infarction (AMI), and explore their role in AMI.

METHODSEnzyme-linked immunoassay (ELISA) was employed for measuring the levels of VEGF and SDF-1 in AMI patients on days 1, 3, 7, 10, and 14 of onset and in normal control subjects. The absolute counts of CD34(+) in the peripheral blood were measured on days 1, 7, and 14 by flow cytometry in AMI patients, with their myocardial enzyme and troponin I detected and electrocardiography (ECG) and echocardiography (UCG) recorded.

RESULTSPeripheral CD34(+) cells obviously increased on day 7 after AMI onset (2.35-/+0.72/microl vs 1.48-/+0.49/micro, P<0.05). VEGF levels were significantly higher in AMI patients than in the control subjects, reaching the peak level and on day 14 (197.56-/+39.87 vs 53.79-/+18.12 pg/ml, P<0.01). SDF-1 level obviously decreased on day 1 after AMI onset (1683.12-/+224.79 vs 2178.67-/+265.34 pg/ml, P<0.01), followed by gradually increased to the control level. Obvious correlation was noted between the level of VEGF on day 7 and the peak level of peripheral CD34(+) cells, and the peak plasma VEGF level was obviously associated with the peak serum CK-MB and troponin I levels.

CONCLUSIONThe stem cells are mobilized into the peripheral blood in the event of AMI. Obviously increased VEGF level following AMI may persist for at least 2 weeks, whereas SDF-1 level undergoes temporary decrement after AMI. The dynamic changes of VEGF and SDF-1 can be related to the mobilization and homing of the stem cells to the injured myocardium.

Adult ; Antigens, CD34 ; blood ; Chemokine CXCL12 ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Humans ; Male ; Myocardial Infarction ; blood ; Time Factors ; Vascular Endothelial Growth Factor A ; blood